Ultracytochemical localization of Na+,K+-activated ATPase in chloride cells from the gills of a euryhaline teleost.

The activity of the electrolyte transport enzyme, sodium, potassium-activated adenosine triphosphatase (Na+,K+-ATPase), in the gills of the pinfish, Lagodon rhomboides, increased markedly following transfer of fish from brackish water to seawater. Cytochemical localization of Na+,K+-ATPase via its potassium-dependent phosphatase (K+-NPPase) activity in the branchial epithelium of pinfish adapted to seawater demonstrated that chloride cells are the major sites for the enzyme. Subcellularly, the heaviest depositions of reaction product were observed lining the cytoplasmic membrane surfaces of the labyrinth of anastomosing plasma membrane tubules that ramifies throughout the chloride cell cytoplasm. Enzyme activity was demonstrated also on the cytoplasmic surface of the apical crypt membrane and on the cytoplasmic surfaces of vesicles in the cytoplasm subjacent to the crypt. Deletion of potassium from the cytochemical incubation medium or inclusion of 10 mM ouabain abolished the reaction products associated with these membranes. The significance of these cytochemical results is discussed with reference to current hypotheses of chloride cell function.